LOCUS       sequence-476853-        3838 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression plasmid encoding chAcK3RS(IPYE) and pylT.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3838)
  AUTHORS   Bryson DI, Fan C, Guo LT, Miller C, Soll D, Liu DR
  TITLE     Continuous directed evolution of aminoacyl-tRNA synthetases.
  JOURNAL   Nat Chem Biol. 2017 Oct 16. doi: 10.1038/nchembio.2474.
  PUBMED    29035361
REFERENCE   2  (bases 1 to 3838)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat Chem 
            Biol. 2017 Oct 16. doi: 10.1038/nchembio.2474."
COMMENT     Sequence Label: pTECH-chAcK3RS(IPYE)
FEATURES             Location/Qualifiers
     source          1..3838
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(join(1937..1942,1943..1959,1960..1965))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     rep_origin      2077..2622
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     2523..2542
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2739..2756
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             3045..3704
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(3111..3130)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 ttgagcgaac gatcaaaaat aagtgccttc ccatcaaaaa aatattctca acataaaaaa
       61 ctttgtgtaa tacttgtaac gctacatgga gattaactca atctagaggg taactagtat
      121 ggataagaag ccgctggatg ttctgatctc tgcgaccggt ctgtggatgt cccgtaccgg
      181 cacgctgcac aagatcaagc actatgagat ttctcgttct aaaatctaca tcgaaatggc
      241 gtgtggtgac catctggttg tgaacaactc tcgttcttgt cgtcccgcac gtgcattccg
      301 ttatcataaa taccgtaaaa cctgcaaacg ttgtcgtgtt tctggtgaag atatcaacaa
      361 cttcctgacc cgttctaccg aaggcaaaac ctctgttaaa gttaaagttg tttctgagcc
      421 gaaagtgaaa aaagcgatgc cgaaatctgt ttctcgtgcg ccgaaaccgc tggaaaatcc
      481 ggtttctgcg aaagcgtcta ccgacacctc tcgttctgtt ccgtctccgg cgaaatctac
      541 cccgaactct ccggttccga cctctgcaag tgcccccgca cttacgaaga gccagactga
      601 caggcttgaa gtcctgttaa acccaaaaga tgagatttcc ctgaattccg gcaagccttt
      661 cagggagctt gagtccgaat tgctctctcg cagaaaaaaa gacctgcagc agatctacgc
      721 ggaagaaagg gagaattatc tggggaaact cgagcgtgaa attaccaggt tctttgtgga
      781 caggggtttt ctggaaataa aatccccgat cctgatccct cttgagtata tcgaaaggat
      841 gggcattgat aatgataccg aactttcaaa acagatcttc agggttgaca agaacttctg
      901 cctgagaccc atgatggctc caaacatttt taactacgct cgcaagcttg acagggccct
      961 gcctgatcca ataaaaattt ttgaaatagg cccatgctac agaaaagagt ccgacggcaa
     1021 agaacacctc gaagagttta ccatgctgaa cttctttcag atgggatcgg gatgcacacg
     1081 ggaaaatctt gaaagcataa ttacggactt cctgaaccac ctgggaattg atttcaagat
     1141 cgtaggcgat tcctgcatgg tctatgggga tacccttgat gtaatgcacg gagacctgga
     1201 actttcctct gcagtagtcg gacccatacc gcttgaccgg gaatggggta ttgataaacc
     1261 ctggataggg gcaggtttcg ggctcgaacg ccttctaaag gttaaacacg actttaaaaa
     1321 tatcaagaga gctgcaaggt ccgagtctta ctataacggg atttctacca acctgtaacc
     1381 taggctgctg ccaccgctga gatctatggc ccgcgcccac aggaaggagc taccggcagc
     1441 ggtgcggact gttgtaactc ataataagaa atgaggccgc tcatggcgtt ctgttgcccg
     1501 tctcgctggt gaaaagaaaa accaccctgg ctaactaagc ggcctgctga ctttctcgcc
     1561 gatcaaaagg cattttgcta ttaagggatt gacgagggcg tatctgcgca gtaagatgcg
     1621 ccccgcattg gaaacctgat catgtagatc gaatggactc taaatccgtt cagccgggtt
     1681 agattcccgg ggtttccgcc aaattcgaaa agcctgctca acgagcaggc ttttttgcat
     1741 ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagatc ccgcaagagg
     1801 cccggcagta ccggcataac caagcctatg cctacagcat ccagggtgac ggtgccgagg
     1861 atgacgatga gcgcattgtt agatttcata cacggtgcct gactgcgtta gcaatttaac
     1921 tgtgataaac taccgcatta aagcttatcg atgataagct gtcaaacatg agaattacaa
     1981 cttatatcgt atggggctga cttcaggtgc tacatttgaa gagataaatt gcactgaaat
     2041 ctagaaatat tttatctgat taataagatg atcttcttga gatcgttttg gtctgcgcgt
     2101 aatctcttgc tctgaaaacg aaaaaaccgc cttgcagggc ggtttttcga aggttctctg
     2161 agctaccaac tctttgaacc gaggtaactg gcttggagga gcgcagccac caaaacttgt
     2221 cctttcagtt tagccttaac cggcgcatga cttcaagact aactcctcta aatcaattac
     2281 cagtggctgc tgccagtggt gcttttgcat gtctttccgg gttggactca agacgatagt
     2341 taccggataa ggcgcagcgg tcggactgaa cggggggttc gtgcatacag tccagcttgg
     2401 agcgaactgc ctacccggaa ctgagtgtca ggcgtggaat gagacaaacg cggccataac
     2461 agcggaatga caccggtaaa ccgaaaggca ggaacaggag agcgcacgag ggagccgcca
     2521 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccaccactg atttgagcgt
     2581 cagatttcgt gatgcttgtc aggggggcgg agcctatgga aaaacggctt tgccgcggcc
     2641 ctctcacttc cctgttaagt atcttcctgg catcttccag gaaatctccg ccccgttcgt
     2701 aagccatttc cgctcgccgc agtcgaacga ccgagcgtag cgagtcagtg agcgaggaag
     2761 cggaatatat cctgtatcac atattctgct gacgcaccgg tgcagccttt tttctcctgc
     2821 cacatgaagc acttcactga caccctcatc agtgccaaca tagtaagcca gtatacactc
     2881 cgctagcgct gatgtccggc ggtgcttttg ccgttacgca ccaccccgtc agtagctgaa
     2941 caggagggac agtaagaggt tccaactttc accataatga aataagatca ctaccgggcg
     3001 tattttttga gttatcgaga ttttcaggag ctaaggaagc taaaatggag aaaaaaatca
     3061 ctggatatac caccgttgat atatcccaat ggcatcgtaa agaacatttt gaggcatttc
     3121 agtcagttgc tcaatgtacc tataaccaga ccgttcagct ggacattacg gcctttttaa
     3181 agaccgtaaa gaaaaataag cacaagtttt atccggcctt tattcacatt cttgcccgcc
     3241 tgatgaatgc tcatccggaa tttcgtatgg caatgaaaga cggtgagctg gtgatatggg
     3301 atagtgttca cccttgttac accgttttcc atgagcaaac tgaaacgttt tcatcgctct
     3361 ggagtgaata ccacgacgat ttccggcagt ttctacacat atattcgcaa gatgtggcgt
     3421 gttacggcga aaacctggcc tatttcccta aagggtttat tgagaatatg tttttcgtct
     3481 cagccaatcc ctgggtgagt ttcaccagtt ttgatttaaa cgtggccaat atggacaact
     3541 tcttcgcccc cgttttcacc atgggcaaat attatacgca aggcgacaag gtgctgatgc
     3601 cgctggcgat tcaggttcat catgccgttt gtgatggctt ccatgtcggc agaatgctta
     3661 atgaattaca acagtactgc gatgagtggc agggcggggc gtaatttttt taaggcagtt
     3721 attggtgccc ttaaacgcct ggttgctacg cctgaataag tgataataag cggatgaatg
     3781 gcagaaattc gaaagcaaat tcgacccggg cggccgcaac ccagcgttcg atgcttct
//
